Figure 5: MiR-424 is repressed transcriptionally by FOXO1.
(a) hASCs were transfected with FOXO1 siRNA for 24 h. siRNA NC (negative control) was used as a negative control. qRT-PCR showed that the knockdown of FOXO1 restored miR-424 expression in hASCs exposed to H2O2 for 24 h. *P < 0.05, ns, not significant. The results indicate the mean ± SD of triplicate experiments. (b) Schematic diagram of the three putative FOXO1 binding sites identified in the miR-424 promoter. (c) A ChIP assay shows that FOXO1 binds to the promoter of miR-424. Three optimal FOXO1 consensus binding sites (BS) in the promoter of miR-424 were identified according to the JASPAR database. Accordingly, we designed three primers containing FOXO1 BS1, BS2 or BS3. Chromatin-bound DNA was immunoprecipitated with the anti-FOXO1 antibody (FOXO1 Ab). IgG was used as a negative control. Thereafter, PCR was performed for the analysis of FOXO1 binding to the promoter of miR-424. The images shown are representative of three independent experiments.
